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Assignment 3- Article Review

Hypothesis

The study aimed to investigate whether the activation of mGgluR2/R3 would result in neuroprotection within a short time after experimental birth asphyxia in 7-day old rats and also whether the antioxidant mechanisms are engaged in the process. The hypothesis, therefore, assumed that the activation of group II mGlu receptors shortly after H-I has a neuroprotective effect, the study wanted to investigate the molecular mechanisms of the effect more closely.

Methods Used

Induction of cerebral hypoxia-ischemia: Neonatal cerebral H-I was induced where rat pups of both sexes were anesthetized with isoflurane in a mixture of nitrous oxide and oxygen. The left common carotid artery was also exposed and cut between double ligatures of silk suture. After 60 min of recovery, the animals were placed in a humidified chamber and exposed to a hypoxic gas for around 75 minutes. The animals were returned to the cages after the hypoxia treatment and housed with their mother at room temperature.

Drug application: Animals were injected intraperitoneally with specific agonists of either mGluR2 (LY379268) or mGluR3, and the side effects monitored.

Evaluation of brain damage: After 14 days, the brains were removed, and both cerebral hemispheres weighed. Brain damage was reflected by a deficit in the wet weight of the ipsilateral (left) ischemic hemisphere and expressed as a percentage of the wet weight of the contralateral (right) control hemisphere.

Determination of ROS level: The ROS levels in the brain hemispheres were determined using 2,7-dichlorofluorescein diacetate.

Determination of glutathione concentration: Glutathione concentrations were measured from the tissues isolated from both left and rights hemispheres after three hours of agonist injection. Reduced glutathione concentrations were determined from supernatants using the Glutathione Assay Kit, which was done in observance of the manufacturer’s procedures.

Determination of antioxidant enzymes activity: Brains were collected after three hours of agonist injection, and then the right and left hemispheres were homogenized separately in an ice-cold 50mM potassium phosphate buffer containing one mM EDTA and centrifuged at 10,000 x g for 15 minutes at four degrees Celsius.

Statistical analysis: The results were expressed as the means ± SEM for each experimental group, where a statistical analysis of the brain damage data was performed via a paired t-test. The remaining data were analyzed through ANOVA.

Conclusion:

The results of the study indicate that the activation of mGluR2 and mGluR3 a short time after H-I triggers neuroprotective mechanisms that act through the inhibition of anti-oxidative stress and ROS production. Comparable results were obtained from the application of both mGluR2 and mGluR3 agonists, an indication that both types of receptors may serve as a potential target for future therapeutic action.

I was convinced with the results since the research was conducted in the right way and involved statistical analysis in obtaining the relationship of mGluR2/R3 agonists in the reduction of oxidative stress and ROS production.